Analysis of mitochondrial DNA protein-coding region in the Yeso Sika deer (Cervus nippon yesoensis)

In the present study, mitochondrial DNA sequences of the Yeso Sika deer (Cervus nippon yesoensis) were studied. Specifically, protein‐coding genes as mitochondrial NADH dehydrogenase subunits (ND1, ND2, ND3, ND4L, ND4, ND5 and ND6), cytochrome c oxidase subunits (CO I and CO III), ATP synthase subunits (ATPase8 and ATPase6) and cytochrome b. Also, phylogenetic analyses on eight mammalian species were performed, including the Muntjac deer (Muntiacus reevesi). The rate of amino‐acid substitution was lowest (3.74%) between Yeso Sika deer and Muntjac deer, and the values between Yeso Sika deer and other species (sheep, cattle, horse, pig, mouse, human and chimpanzee) were 6.63%, 7.30%, 12.55%, 13.03%, 23.59%, 24.82% and 25.04%, respectively. Among them, the highest value of divergence was recognized in ATPase8, and the second structure of ATPase8 showed a difference between the Yeso Sika deer and Muntjac deer as a result of the substitution of 34His→Tyr and 49Thr→Ile. In addition, we identified a substitution of an amino‐acid sequence (19Thr→Ala) between the Yeso Sika deer and Yakushima Sika deer (C. n. yakushimae). From these results, ATPase8 was also a variable region in Cervidae.     

Conformational change in hamster scrapie prion protein (PrP27-30) associated with proteinase K resistance and prion infectivity

The scrapie prion protein (PrP27-30) is a crucial component of the prion and is responsible for its transmissibility. Structural information on this protein is limited because it is insoluble and shows aggregated properties. In this study, PrP27-30 was effectively dispersed using sonication under the weak alkaline condition. Subsequently, the small PrP27-30 aggregates were subjected to different pH, heat, and denaturing conditions. The loss of proteinase K (PK) resistance of PrP27-30 and prion infectivity were monitored along with spectroscopic changes. Prion inactivation could not be achieved by the loss of PK resistance alone; a significant loss of the PrP27-30 amyloid structure, which was represented by a decrease in thioflavin T fluorescence, was required for the loss of transmissibility.     

In vitro differentiation of canine celiac adipose tissue-derived stromal cells into neuronal cells

To investigate in vitro differentiation of canine adipose tissue-derived stromal cells (ATSCs) into neuronal cells, ATSCs from celiac adipose tissue in clinically healthy beagle dogs were treated with 100 muM dibutyryl cyclic adenosine monophosphate (dbcAMP) and 125 muM isobuthylmethylxanthine (IBMX). ATSCs were morphologically changed into differentiated ATSCs from spindle-shaped cells to neuron-like cells with numerous processes after the treatment. Expression of neuron-specific enolase (NSE) as an early neuron specific marker protein was detected in both ATSCs and differentiated ATSCs, however diachronic increase of NSE expression was observed in differentiated ATSCs after the treatment with dbcAMP/IBMX. In addition, neurofilament-68 (NF-68) as an early to mature neuron specific marker protein was weakly expressed in differentiated ATSCs. Neuron specific glutamate and glucose transporter (EAAC1 and GLUT-3, respectively) mRNAs were strongly expressed in differentiated ATSCs compared with those in ATSCs, although glia specific glutamate transporter mRNA (GLT-1) was also detected in differentiated ATSCs. ATSCs can differentiate into early to mature neuronal cells and are candidate cells for autologous nerve regeneration therapy, although additional research is needed to examine functional characteristics of differentiated ATSCs.     

Kazusa Marker DataBase: a database for genomics,genetics, and molecular breeding in plants

In order to provide useful genomic information for agronomical plants, we have established a database, the Kazusa Marker DataBase (http://marker.kazusa.or.jp). This database includes information on DNA markers, e.g., SSR and SNP markers, genetic linkage maps, and physical maps, that were developed at the Kazusa DNA Research Institute. Keyword searches for the markers, sequence data used for marker development, and experimental conditions are also available through this database. Currently, 10 plant species have been targeted: tomato (Solanum lycopersicum), pepper (Capsicum annuum), strawberry (Fragaria × ananassa), radish (Raphanus sativus), Lotus japonicus, soybean (Glycine max), peanut (Arachis hypogaea), red clover (Trifolium pratense), white clover (Trifolium repens), and eucalyptus (Eucalyptus camaldulensis). In addition, the number of plant species registered in this database will be increased as our research progresses. The Kazusa Marker DataBase will be a useful tool for both basic and applied sciences, such as genomics, genetics, and molecular breeding in crops.     

Relationships between nutrients and sucrose concentrations in sugarcane juice and use of juice analysis for nutrient diagnosis in Japan

Sugarcane is an important economic crop in southwest Japan, but its production is decreasing. To increase sugar production, both sugarcane yield and quality should be improved. Fertilizer management is one of the factors that influence sugarcane quality. We accordingly focused on nutrients present in sugarcane juice and attempted to identify the key factors affecting sugarcane quality. We collected sugarcane samples from 2013 to 2015 from all of the sugar mills in Japan and examined the relationships between juice nutrients and sucrose concentration. Juice analysis over 3 year showed that potassium (K⁺) and chloride (Cl^?) were the most abundant cation and anion in the juice and that both negatively correlated with the sucrose concentration. K⁺ and Cl^? concentrations significantly varied depending on production areas and those with higher K⁺ and Cl^? concentrations had a low sucrose concentration. This finding suggests that sugarcane in those areas may have been supplied with these two ions in excess. Electrical conductivity (EC) in the juice always positively correlated with K⁺ and Cl^? concentrations. EC may thus be a reliable indicator of K⁺ and Cl^? concentrations and could be used for nutrient diagnosis because of its ease of measurement. For improving sugarcane quality, we recommend that potassium chloride, which supplies both K⁺ and Cl^? and is a commonly used potassium fertilizer for sugarcane production in Japan, should be used in lower quantities in a year following one in which the EC of sugarcane juice at harvest is found to be high.     

Strength properties of glued laminated timber made from edge-glued laminae I: strength properties of edge-glued karamatsu (Larix kaempferi) laminae

The object of this study was to investigate the strength properties of edge-glued laminae and to propose a suitable grading method based on the lamina modulus of elasticity (MOE). Edge-glued laminae composed of lumber with similar MOEs (uniform laminae) and edge-glued laminae produced by randomly gluing lumber independent of MOE (random laminae) were made from karamatsu (Larix kaempferi) lumber having the same thickness and length, but various widths. For both the uniform and random laminae, there was a strong correlation between MOE values measured using the longitudinal vibration technique, the static bending test, and a grading machine. The average values of bending, tensile, and compressive strengths of the uniform laminae were similar to those of the random laminae. On the other hand, the average strength of laminae without end joints was significantly higher than that of finger-jointed laminae for both uniform and random laminae. Finger-joints and knots played a significant role in the failure of specimens, but the edge-gluing and the difference in MOE within an edge-glued lamina did not appear to affect the strength properties. The bending, tensile, and compressive strengths of edge-glued laminae were strongly correlated to the lamina MOE.     

Effect of lodging on the level of mycotoxins in wheat,barley, and rice infected with the <Emphasis Type="Italic">Fusarium graminearum</Emphasis> species complex

Lodging is one possible risk factor that leads to increased cereal mycotoxin contamination, but few reports have been published on the subject. We examined the effects of lodging on the level of deoxynivalenol (DON) and nivalenol (NIV) contamination in wheat, barley, and rice infected with the Fusarium graminearum species complex. Case-control and intervention studies were applied to test the hypothesis that lodging increases the level of mycotoxin contamination. A total of 66 grain samples were collected from each field in 12 Japanese prefectures from 2002 to 2006. Each sample set consisted of grains from lodged and nonlodged plants. The concentration of DON + NIV in lodged plants was significantly higher than in nonlodged plants. All samples of wheat and barley were contaminated with DON and NIV; however, most of the lodged rice samples were contaminated only with NIV. In intervention trials to investigate the effects of lodging duration, a small area of wheat inoculated with the pathogen was completely lodged by trampling. Even with 5 days of lodging, the levels of DON + NIV in wheat grain at harvest increased by 27–51% compared to nonlodged control plots. For rice, half of each plot area was completely lodged by trampling 20 days before harvest. The level of NIV in lodged rice grain was significantly higher than that in nonlodged rice at optimum and delayed harvests, because lodging significantly increased the level of Fusarium mycotoxins in the three crops. Thus, practices (e.g., rational use of fertilizers) to avoid lodging should reduce the risk of mycotoxin contamination. This is the first epidemiological study on the effect of lodging on mycotoxin production by the F. graminearum species complex in wheat, barley, and rice.     

Different cross-reactivity of human and rodent sera to Tula virus and Puumala virus

Tula virus (TULV) and Puumala virus (PUUV) are hantaviruses carried by the bank vole (Myodes glareolus) and European common vole (Microtus arvalis), respectively. PUUV is a causative agent of hemorrhagic fever with renal syndrome (HFRS), while TULV is thought to be apathogenic to humans. The N-terminal regions of the N proteins from TULV and PUUV were expressed and applied as enzyme-linked immunosorbent assay (ELISA) antigens. Colonized Japanese grass voles (Microtus montebelli) and BALB/c mice were used for experimental inoculation of the vole-borne hantaviruses TULV and PUUV. Voles and mice showed significant antibody production toward both viruses, but these antisera showed little cross-reactivity between TULV and PUUV in the immunofluorescence antibody assay and ELISA. In contrast, sera from patients with HFRS caused by PUUV exhibited high cross-reactivity against the TULV antigen, and sera from a natural rodent reservoir showed moderate cross-reactivity against the heterologous antigen, indicating that the antigenic cross-reactivity between TULV and PUUV differs in sera from rodents and humans.     

Severe calcification of mucocutaneous and gastrointestinal tissues induced by high dose administration of vitamin D in a puppy

A three-month-old male Bull Terrier was referred to the Animal Medical Centre, Nihon University with chief complaints of subacute emesis and lethargy. Severe leukocytosis, high CRP, hypercalcemia and hypochloremia were detected. Moreover, severe calcification of gingival mucosa and abdominal skin, and abnormalities of the skeletal system were discerned. Abdominal X-ray and endoscopic examination revealed ulcer and hemorrhage on the mucosal membrane of the stomach. This might have been due to injections of high dose vitamin D at 3 and 2 weeks ago by another practioner, according to the detailed history of medication. After two months, a gastrointestinal and skin disorder disappeared, although calcification of the stomach membranes remained and abnormality of the skeletal system had worsened. Therefore, vitamin D should be carefully administrated to a puppy.